- Volume 55, Issue 5, 2005

A novel bacterial strain, MC10T, was isolated from a compost sample produced in a composting machine utilizing soil from Motobu-town, Okinawa, Japan. The isolate was Gram-negative, but produced endospores. These conflicting characters prompted a taxonomic study of the isolate. The isolate was examined using a combination of phenotypic characterization, cellular fatty acid analysis, DNA base composition determination and 16S rRNA gene sequence analysis. Phylogenetic analysis, based on 16S rRNA gene sequence comparisons, placed strain MC10T within the genus Paenibacillus. As in other Paenibacillus species, the isolate contained anteiso-C15 : 0 as the major fatty acid and the DNA G+C content was 47·0 mol%. However, 16S rRNA gene sequence similarity values of less than 95·6 % were found between this isolate and all members of the genus Paenibacillus. Based upon these results, strain MC10T (=GTC 1835T=JCM 12774T=CCUG 50090T) should be designated as the type strain of a novel species within the genus Paenibacillus, Paenibacillus motobuensis sp. nov.

A Gram-variable, motile, endospore-forming, slightly halophilic bacterial strain, DSW-10T, was isolated from Dokdo, an island located at the edge of the East Sea, Korea, and was characterized taxonomically by using a polyphasic approach. This isolate grew optimally at 37 °C and in the presence of 4–5 % NaCl. Strain DSW-10T had cell-wall peptidoglycan based on meso-diaminopimelic acid, MK-7 as the predominant menaquinone, and anteiso-C15 : 0, iso-C15 : 0, anteiso-C17 : 0 and iso-C16 : 0 as major fatty acids. Major polar lipids were diphosphatidylglycerol, phosphatidylglycerol, phosphatidylethanolamine and unidentified phospholipids. The DNA G+C content was 36·7 mol%. Phylogenetic analyses based on 16S rRNA gene sequences showed that strain DSW-10T is phylogenetically affiliated to the genus Virgibacillus, and exhibited sequence similarity of 95·3–98·7 % to the type strains of Virgibacillus species. DNA–DNA relatedness levels between strain DSW-10T and the type strains of some phylogenetically related Virgibacillus species were in the range 8·4–17·5 %. On the basis of phenotypic properties and phylogenetic and genetic distinctiveness, strain DSW-10T (=KCTC 3933T=DSM 16826T) was classified as the type strain of a novel Virgibacillus species, for which the name Virgibacillus dokdonensis sp. nov. is proposed.

A Gram-positive, non-motile actinobacterium, designated strain YIM 70177T, was isolated from a saline soil sample from Qinghai Province, north-west China. Phylogenetic analysis of this organism based on 16S rRNA gene sequences revealed 98·3 % similarity to Isoptericola variabilis DSM 10177T. Chemotaxonomic data determined for the isolate, such as the peptidoglycan type A4α, variation l-lys–d-Asp, supported the placement of strain YIM 70177T within the genus Isoptericola. Galactose was detected as cell wall sugar. Phospholipids identified were phosphatidylinositol, phosphatidylglycerol and diphosphatidylglycerol, and the predominant menaquinones were MK-9(H4) and MK-9(H2). The major fatty acids were ai-C15 : 0, C16 : 0, ai-C17 : 0 and i-C15 : 0. The DNA G+C content was 72·8 mol%. The low level of DNA–DNA relatedness (27·4 %) to I. variabilis DSM 10177T in combination with differentiating chemotaxonomic and physiological data demonstrated that the isolate YIM 70177T should be classified as representing a novel species of the genus Isoptericola. The name Isoptericola halotolerans sp. nov. is proposed, with strain YIM 70177T (=DSM 16376T=KCTC 19046T) as the type strain.

The taxonomic positions of two clinically isolated actinomycetes were established using a polyphasic approach. The two strains, IFM 10032T, isolated from ear discharge of a 28-year-old Japanese female patient with external otitis, and IFM 10148, isolated from pleural fluid of a 60-year-old Japanese male patient with bronchitis, possessed meso-diaminopimelic acid as the diagnostic amino acid, MK-9(H2) as the predominant menaquinone and mycolic acids ranging from 58 to 64 carbons. The 16S rRNA gene sequences of the two strains were most closely related to those of Gordonia aichiensis, Gordonia sputi and ‘Gordonia jacobaea’. Differences in several phenotypic characteristics together with genotypic distinctiveness distinguish strains IFM 10032T and IFM 10148 from these three species. DNA–DNA hybridization results and the combination of genotypic and phenotypic data showed that the two strains belong to a single species, and merit recognition of a novel species within the genus Gordonia. The name proposed for this taxon is Gordonia otitidis sp. nov.; the type strain is IFM 10032T (=DSM 44809T=JCM 12355T=NBRC 100426T).

A spore-forming, halophilic bacterium, designated strain BH163T, was isolated from a salt lake in China. Cells were motile, strictly aerobic rods that contained type A1γ peptidoglycan with meso-diaminopimelic acid as the diagnostic diamino acid. The isolate showed Gram- and catalase-positive reactions and formed a terminal endospore with a swollen sporangium. The major cellular fatty acids were anteiso-C15 : 0, iso-C15 : 0, anteiso-C17 : 0 and iso-C16 : 0. The genomic DNA G+C content of the strain was 41·0 mol%. Comparative analysis of 16S rRNA gene sequences showed that strain BH163T formed a distinct line within the phyletic group classically defined as the genus Bacillus and was most closely related to the taxa [Bacillus] haloalkaliphilus DSM 5271T and Filobacillus milosensis DSM 13259T, with 16S rRNA gene sequence similarities of 95·9 and 94·5 %, respectively. On the basis of physiological and molecular properties, it is proposed that [Bacillus] haloalkaliphilus DSM 5271T is reclassified in the new genus Alkalibacillus as Alkalibacillus haloalkaliphilus gen. nov., comb. nov. Strain BH163T (=KCTC 3916T=DSM 16460T) was assigned as the type strain of the novel species Alkalibacillus salilacus.

An actinomycete, strain Z33T, was isolated from a cyanobacterial biofilm in the Cave of Bats, near Zuheros (Cordoba, southern Spain). 16S rRNA gene sequence analysis showed that strain Z33T formed a distinct phyletic line within the genus Agromyces. This isolate could be readily distinguished from representatives of all recognized Agromyces species on the basis of a broad range of phenotypic characteristics and DNA–DNA relatedness data. Genotypic and phenotypic properties indicate that strain Z33T represents a novel species, for which the name Agromyces subbeticus sp. nov. is proposed. The type strain is Z33T (=HKI 0340T=DSM 16689T=NCIMB 14025T).

The taxonomic positions of two soil actinomycetes, strains 117T and 43401T, provisionally assigned to the genus Nocardia were determined in a polyphasic study. The organisms were found to have phenotypic properties typical of members of the genus Nocardia, and formed a distinct branch in the Nocardia 16S rRNA gene tree. It was evident from the phylogenetic data that the isolates were most closely, albeit loosely, associated with the type strains of Nocardia nova and Nocardia pseudobrasiliensis. However, all four of these organisms were readily distinguishable from one another using DNA–DNA relatedness and phenotypic data. It is evident from the genotypic and phenotypic data that the two isolates should be recognized as novel species of the genus Nocardia. It is proposed, therefore, that strains 117T (=CGMCC 4.1904T=JCM 12860T) and 43401T (=CGMCC 4.1905T=JCM 12861T) be classified in the genus Nocardia as the type strains of Nocardia miyunensis sp. nov. and Nocardia jiangxiensis sp. nov., respectively.

A coccoid actinobacterium strain (designated YIM 70178T) was isolated from a soil sample collected in Qinghai Province, China. The isolate grew well with an optimum salt concentration of 10–15 % (KCl, w/v) but scarcely or not at all without salt. The cell-wall peptidoglycan type was A4α, l-lys–gly–l-Glu. The major polar lipids were diphosphatidylglycerol, phosphatidylglycerol, an unknown phospholipid and an unknown glycolipid. The predominant menaquinones were MK-8 and MK-9. The major fatty acid was anteiso-C15 : 0. The DNA G+C content was 57·9 mol%. Phylogenetic analysis based on 16S rRNA gene sequences showed that strain YIM 70178T was most closely related to the type strain of Yania halotolerans. DNA–DNA hybridization and comparison of physiological and chemotaxonomic characteristics demonstrated that strain YIM 70178T was different from Yania halotolerans. The name Yania flava sp. nov. is proposed, with strain YIM 70178T (=DSM 16377T=KCTC 19047T) as the type strain. Based on the phenotypic characteristics and phylogenetic position, as determined by 16S rRNA gene analysis and 16S rRNA signature nucleotide data, the genus description of Yania is therefore emended and strains YIM 70085T and YIM 70178T represent a novel family of the suborder Micrococcineae, for which the name Yaniaceae fam. nov. is proposed.

An actinomycete, strain FXJ46T, was isolated from cypress forest soil in northern China and shown to have chemotaxonomic and morphological properties consistent with streptomycetes. It developed greyish aerial mycelium and pinkish-brown substrate mycelium on oatmeal agar. Phylogenetic analyses based on an almost complete 16S rRNA gene sequence of the strain and on the 120 nucleotide variable γ-region of this molecule showed that it formed a distinct (but closely associated) line with Streptomyces griseoaurantiacus DSM 40430T in Streptomyces trees. However, the DNA–DNA relatedness between the two strains was only 48·8 %. A number of phenotypic properties also readily distinguished the isolate from S. griseoaurantiacus and related Streptomyces species with validly published names. It is proposed, therefore, that this organism be classified as a novel species of the genus Streptomyces, for which the name Streptomyces jietaisiensis sp. nov. is proposed. The type strain is FXJ46T (=AS 4.1859T=JCM 12279T).

A Gram-variable, motile, endospore-forming, slightly halophilic bacterial strain, designated SW-211T, was isolated from a tidal flat of the Yellow Sea in Korea, and was characterized taxonomically by using a polyphasic approach. The organism grew optimally at 37 °C and in the presence of 2–3 % NaCl. Comparative 16S rRNA gene sequence analysis showed that strain SW-211T forms a distinct phylogenetic lineage within the radiation of the cluster comprising Bacillus species. Strain SW-211T had cell-wall peptidoglycan based on meso-diaminopimelic acid. The predominant menaquinone was MK-7 and the major fatty acids were anteiso-C15 : 0 (34·8 %), iso-C15 : 0 (15·6 %), iso-C16 : 0 (12·5 %) and iso-C14 : 0 (10·0 %). The DNA G+C content was 35·2 mol%. Strain SW-211T exhibited levels of 16S rRNA gene sequence similarity of <96·2 % with respect to the type strains of recognized Bacillus species. On the basis of its phenotypic properties and phylogenetic distinctiveness, strain SW-211T (=KCTC 3898T=DSM 16303T) was classified in the genus Bacillus as the type strain of a novel species, for which the name Bacillus litoralis sp. nov. is proposed.

Eight coagulase-negative, oxidase-negative and novobiocin-susceptible staphylococcal strains were isolated from the gastrointestinal tracts of South American squirrel monkeys (Saimiri sciureus L.). These strains were differentiated from known staphylococcal species on the basis of 16S rRNA gene and hsp60 gene sequencing, and from the most closely related species by using DNA–DNA hybridization, ribotyping, whole-cell protein profiles and biotyping. Phylogenetic analysis based on 16S rRNA gene sequences confirmed that these strains are members of the Staphylococcus aureus species group (99 % similarity) but are biochemically similar to Staphylococcus piscifermentans, from which they can be phenotypically distinguished by resistance to polymyxin B, acid production from d-mannitol, the inability to hydrolyse aesculin and DNA and the absence of α-glucosidase. On the basis of these analyses, a novel species of the genus Staphylococcus is described, for which the name Staphylococcus simiae sp. nov. is proposed, with CCM 7213T (=LMG 22723T) as the type strain.

The taxonomic position of an actinomycete strain isolated from soil from Natore, Bangladesh, was examined by using a polyphasic approach. The strain, designated AAB-4T, was assigned to the genus Streptomyces on the basis of chemical and morphological criteria. It formed Rectiflexibiles aerial hyphae that carried long chains of rounded spores. The 16S rRNA gene of strain AAB-4T was sequenced directly and then compared with those of previously studied streptomycetes following the generation of two phylogenetic trees by using maximum-likelihood and neighbour-joining algorithms. This confirmed the assignment of the novel strain to the genus Streptomyces. This strain showed a high level of 16S rRNA gene sequence similarity to Streptomyces thermoviolaceus, Streptomyces thermodiastaticus and Streptomyces longisporus, among others, but could be distinguished from them by phenotypic and physiological traits. This micro-organism produces bis-(2-ethylhexyl)phthalate, an antibacterial and antifungal agent. It is proposed that strain AAB-4T be classified as a novel species within the genus Streptomyces, as Streptomyces bangladeshensis sp. nov. (type strain, AAB-4T=LMG 22738T=NRRL B-24326T).

A polyphasic study was undertaken to clarify the taxonomic position of Nonomuraea flexuosa DSM 41386T. The distinct 16S rRNA gene sequence phyletic branch formed by this strain was equated with nine related monophyletic clades composed of representatives of the genera classified in the family Streptosporangiaceae. The organism produced a PCR product characteristic of this taxon when examined using a set of oligonucleotide primers specific for members of the family Streptosporangiaceae. Strain DSM 41386T could also be distinguished from representatives of the nine genera assigned to this family using a combination of chemotaxonomic, morphological and physiological properties. It is evident from the genotypic and phenotypic data that strain DSM 41386T is misclassified in the genus Nonomuraea and merits recognition as a monospecific genus within the family Streptosporangiaceae. It is proposed that the name Thermopolyspora flexuosa gen. nov., comb. nov., nom. rev. be used for this purpose, with the type strain DSM 41386T (=NRRL B-24348T).

Two moderately halophilic spore-forming bacteria were isolated from salt lakes in the Xinjiang region of China. The two strains, designated AD-6T and D-8T, were aerobic, Gram-positive, rod-shaped and motile by means of peritrichous flagella. Strains AD-6T and D-8T grew in the presence of 0·5–20 % and 0·5–25 % (w/v) NaCl in complex medium, respectively. Their cell-wall peptidoglycan was of the l-orn–d-Asp type. The major menaquinone found in both strains was menaquinone-7 (MK-7). The fatty acid profile contained a large amount of branched fatty acids; the main fatty acids were anteiso-C15 : 0, anteiso-C17 : 0, iso-C15 : 0 and iso-C16 : 0. The DNA G+C content of strains D-8T and AD-6T was 41·4 and 42·2 mol%, respectively. Phylogenetic analysis based on 16S rRNA gene sequences revealed that strains D-8T and AD-6T were located in the genus Halobacillus. Levels of 16S rRNA gene sequence similarity between the isolated strains and the type strains of Halobacillus species were in the range 96·2–99·5 %. DNA–DNA relatedness values of 17·0–52·2 % were found between the two strains and other Halobacillus species. The DNA–DNA relatedness value between D-8T and AD-6T was 50·6 %. On the basis of phenotypic and chemotaxonomic properties, phylogenetic analysis and genomic distinctiveness, strains D-8T and AD-6T should be placed in the genus Halobacillus as two novel species, for which the names Halobacillus dabanensis sp. nov. (type strain=JCM 12772T=CGMCC 1.3704T) and Halobacillus aidingensis sp. nov. (type strain=JCM 12771T=CGMCC 1.3703T) are proposed, respectively.

The taxonomic position of an actinomycete isolated from a brown hypermagnesian ultramafic soil was examined using a polyphasic approach. The organism, which was designated SBHS Strp1T, was found to have chemical and morphological properties typical of Amycolatopsis strains. It was most closely associated with Amycolatopsis kentuckyensis, Amycolatopsis lexingtonensis, Amycolatopsis rifamycinica, Amycolatopsis pretoriensis and Amycolatopsis tolypomycina on the basis of 16S rRNA gene sequence data, and showed a unique pattern of phenotypic properties that distinguished it from the type strains of these taxa. The combined genotypic and phenotypic data show that the organism merits description as a novel species of Amycolatopsis. The name proposed for the novel species is Amycolatopsis plumensis sp. nov.; the type strain is SBHS Strp1T (=DSM 44776T=NRRL B-24324T).

A novel anaerobic, thermophilic, alkalitolerant bacterium, strain 2204T, was isolated from a hot spring of the Baikal Lake region. The cells of strain 2204T were straight rods of variable length, Gram-positive with an S-layer, motile with one to two lateral flagella, and often formed aggregates of 3–15 cells. The isolate was shown to be an obligate anaerobe oxidizing CO and producing equimolar quantities of H2 and CO2 according to the equation CO+H2O→CO2+H2. No organic substrates were used as energy sources. For lithotrophic growth on CO, 0·2 g acetate or yeast extract l−1 was required but did not support growth in the absence of CO. Growth was observed in the temperature range 37–68 °C, the optimum being 55 °C. The pH range for growth was 6·7–9·5, the optimum pH being 8·0. The generation time under optimal conditions was 1·3 h. The DNA G+C content was 45 mol%. Penicillin, erythromycin, streptomycin, rifampicin, vancomycin and tetracycline completely inhibited both growth and CO utilization by strain 2204T. Thus, isolate 2204T was found to be the first known moderately thermophilic and alkalitolerant H2-producing anaerobic carboxydotroph. The novel bacterium fell within the cluster of the family Peptococcaceae within the low-G+C-content Gram-positive bacteria, where it formed a separate branch. On the basis of morphological, physiological and phylogenetic features, strain 2204T should be assigned to a novel genus and species, for which the name Thermincola carboxydiphila gen. nov., sp. nov. is proposed. The type strain is strain 2204T (=DSM 17129T=VKM B-2283T=JCM 13258T).

A novel xylan-degrading bacterium, S3-ET, was isolated from the biofilm of a membrane bioreactor. The cells of this strain were Gram-positive, non-motile, non-spore-forming rods, produced primary branches and formed yellow colonies on nutrient agar. The strain had chemotaxonomic markers that were consistent with classification in the genus Microbacterium, i.e. MK-12, MK-11 and MK-13 as the major menaquinones, predominant iso- and anteiso-branched cellular fatty acids, glucose and galactose as the cell-wall sugars, peptidoglycan-type B2β with glycolyl residues and a DNA G+C content of 69·7 mol%. Phylogenetic analysis, based on 16S rRNA gene sequencing, showed that strain S3-ET is most similar to Microbacterium hominis IFO 15708T and Microbacterium foliorum DSM 12966T (97·6 and 97·4 % sequence similarity, respectively), and that it forms a separate lineage with M. hominis in the genus Microbacterium. DNA–DNA hybridization results and phenotypic properties showed that strain S3-ET could be distinguished from all known Microbacterium species and represented a novel species, for which the name Microbacterium xylanilyticum sp. nov. is proposed; the type strain is S3-ET (=DSM 16914T=KCTC 19079T).

Two actinomycete strains, IFM 0354T and IFM 0576, isolated from Japanese patients, were found to have morphological, biochemical and chemotaxonomic properties consistent with their classification in the genus Nocardia. The strains resembled Nocardia otitidiscaviarum and Nocardia uniformis in their phenotypic properties. Comparative 16S rRNA gene sequence analysis showed that the strains are closely related to Nocardia seriolae. DNA–DNA relatedness values and phenotypic differences from N. seriolae indicated that the strains belong to a novel species of Nocardia, for which the name Nocardia concava sp. nov. is proposed. The type strain is IFM 0354T (=NBRC 100430T=JCM 12351T=DSM 44804T).

A novel solvent-producing, anaerobic clostridium, strain P7T, was isolated from sediment from an agricultural settling lagoon after enrichment with CO as the substrate. The metabolism of this Gram-positive, motile, spore-forming rod was primarily acetogenic. Acetate, ethanol, butyrate and butanol were the end-products of metabolism. Strain P7T grew on CO, H2/CO2, glucose, galactose, fructose, xylose, mannose, cellobiose, trehalose, cellulose, starch, pectin, citrate, glycerol, ethanol, propanol, 2-propanol, butanol, glutamate, aspartate, alanine, histidine, asparagine, serine, betaine, choline and syringate as sole substrates. Growth was not supported by methanol, formate, d-arabinose, fucose, lactose, melibiose, amygdalin, gluconate, lactate, malate, arginine, glutamine or vanillate. Nitrate reduction, production of indole, gelatin hydrolysis and aesculin hydrolysis were not observed. Analysis of the 16S rRNA gene sequence of the isolate showed that it was closely related to Clostridium scatologenes ATCC 25775T (99·7 % sequence similarity) and clostridial strain SL1T (99·8 % sequence similarity). Strain SL1 had been classified as a strain of C. scatologenes. However, DNA–DNA reassociation analysis showed that both strain P7T and strain SL1 represented novel clostridial species. It is proposed that strain P7T (=ATCC BAA-624T=DSM 15243T) be classified as the type strain of Clostridium carboxidivorans sp. nov. and that strain SL1T (=ATCC BAA-623T=DSM 12750T) be reclassified as the type strain of Clostridium drakei sp. nov.